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題名 | 利用PCR-RFLP技術快速鑑定臺灣進口農產品中常見的介殼蟲(同翅目:介殼蟲總科)=Application of the PCR-RFLP Technique for the Rapid Diagnosis of Scale Insects (Homoptera: Coccoidea) on Imported Agricultural Products in Taiwan |
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作者 | 邱一中; 吳文哲; 翁振宇; 陳淑佩; 石正人; | 書刊名 | 臺灣昆蟲 |
卷期 | 24:2 2004.06[民93.06] |
頁次 | 頁159-171 |
分類號 | 385.752 |
關鍵詞 | 介殼蟲; 快速鑑定; 限制酶片段長度多態型; 分子檢索表; Scale insect; Rapid identification; Restriction fragment length polymorphism; RFLP; Molecular key; |
語文 | 中文(Chinese) |
中文摘要 | 依據海關人員截獲農產品上附有的介殼蟲,經鑑定共計有30種,本研究針對其中16種經常截獲的介殼蟲,建立以核糖體形成基因為基礎的聚合酶連鎖反應-限制酶片段長度多態型技術(ribosomal DNA / polymerase chain reaction-restriction fragment length polymorphism, rDNA/PCR-RFLP),做為介殼蟲分子快速診斷鑑定的操作標準流程,提供海關、機場的檢疫人員依操作標準進行常見介殼蟲的診斷鑑定。試驗篩選出NS7和ITS6引子對,可供增幅供試的16種介殼蟲之rDNA/ITS1特定區域。並進一步篩選出MspI、TaqI和HaeIII核酸內切酶的酶切圖譜具有較高的種類特異性,可作為種類鑑定依據。此外,利用分類學慣用的檢索表概念,以RFLP酶切圖譜差異之分生特徵,建立試驗16種介殼蟲的分子檢索表,提供快速、簡單且準確的診斷鑑定方式。 |
英文摘要 | There are 30 scale insect species which have been intercepted from agricultural products coming into Taiwan by quarantine workers at customs. This study attempted to develop a molecular diagnostic technique (polymerase chain reaction-restriction fragment length polymorphism, PCR-RFLP) using genomic DNA markers (ribosomal DNA, rDNA) for identifying 16 scale insect species. NS7 and ITS6 primer sets were used to amplify ITS1 and its flanking regions of rDNA from genomic DNA as a template extracted from a single specimen. DNA extracted from these 16 scale insect species yielded a single fragment after PCR amplification. These PCR products were then cut with various restriction endonucleases in order to compare the results of restriction fragment length polymorphism. Our results also reveal that it is possible to discriminate these 16 scale insect species based on the species-specific patterns acquired from digesting the PCR products with the endonucleases, TaqI, MspI, and HaeIII. Futhermore, we also a set up a molecular key for these 16 scale insect species based on their RFLP digestion patterns. |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。